Applications

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application

Abreviews

Notes

IP

Use a concentration of 5 µg/ml.

WB

Use a concentration of 1 µg/ml. Detects a band of approximately 105 kDa (predicted molecular weight: 43 kDa).

Target

FunctionA SLe(x)-type proteoglycan, which through high affinity, calcium-dependent interactions with E-, P- and L-selectins, mediates rapid rolling of leukocytes over vascular surfaces during the initial steps in inflammation. Critical for the initial leukocyte capture.(Microbial infection) Acts as a receptor for enterovirus 71.

Tissue specificityExpressed on neutrophils, monocytes and most lymphocytes.

Post-translationalmodificationsDisplays complex, core-2, sialylated and fucosylated O-linked oligosaccharides, at least some of which appear to contain poly-N-acetyllactosamine with varying degrees of substitution. Mainly disialylated or neutral forms of the core-2 tetrasaccharide, Galbeta1-->4GlcNAcbeta1-->6(Galbeta1-->3)GalNAcOH. The GlcN:GalN ratio is approximately 2:1 and the Man:Fuc ratio 3:5. Contains about 14% fucose with alpha-1,3 linkage present in two forms: One species is a disialylated, monofucosylated glycan, and the other, a monosialylated, trifucosylated glycan with a polylactosamine backbone. The fucosylated forms carry the Lewis antigen and are important for interaction with selectins and for functioning in leukocyte rolling. The modification containing the sialyl Lewis X glycan is on Thr-57. No sulfated O-glycans. Some N-glycosylation.Sulfation, in conjunction with the SLe(x)-containing glycan, is necessary for P- and L-selectin binding. High affinity P-selectin binding has a preferred requirement for the isomer sulfated on both Tyr-48 and Tyr-51, whereas L-selectin binding requires predominantly sulfation on Tyr-51 with sulfation on Tyr-48 playing only a minor role. These sulfations play an important role in L- and P-selectin-mediated neutrophil recruitment, and leukocyte rolling.

Anti-CD162 antibody images

CD162 was immunoprecipitated using 0.5mg Rat thymus tissue extract, 5µg of Rabbit polyclonal to CD162 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

The antibody was incubated under agitation with Protein G beads for 10min, Rat thymus tissue extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab66882.

Predicted band size : 43 kDaObserved band size : 105 kDa (why is the actual band size different from the predicted?)Additional bands at : 200 kDa. We are unsure as to the identity of these extra bands.CD162 contains a number of potential glycosylation and phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

References for Anti-CD162 antibody (ab66882)

ab66882
has not yet been referenced specifically in any publications.

Publishing research using ab66882? Please let us know so that we can cite the reference in this datasheet.